Bio-Rad Mini-PROTEAN® Tetra Cell - Manual (Page 27)

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5. 10x electrode (running) buffer, pH 8.3
30.3 g Tris base (15 g/L)
144.1 g glycine (72 g/L)
Bring total volume up to 1,000 ml with deionized water. Do not
adjust pH. Alternatively, electrophoresis running buffer 10x Tris/
glycine, 1 L (Bio-Rad catalog #161-0734) can be used.
Usage: Dilute 100 ml of 10x stock with 900 ml deionized water
for each electrophoresis run.
Gel Formulations (10 ml)
1. Prepare the monomer solution by mixing all reagents except
the TEMED and 10% APS. Degas the mixture for 15 min.
30 % Degassed
Percent DDI H
2
O Acrylamide/Bis Gel buffer
gel (ml) (ml) (ml)
4% 6.2 1.3 2.5
5% 5.8 1.7 2.5
6% 5.5 2.0 2.5
7% 5.2 2.3 2.5
8% 4.8 2.7 2.5
9% 4.5 3.0 2.5
10% 4.2 3.3 2.5
*Resolving Gel Buffer – 1.5 M Tris-HCl, pH 8.8
*Stacking Gel Buffer – 0.5 M Tris-HCl, pH
2. Immediately prior to pouring the gel, add:
50 ml APS and TEMED (5 µl for resolving gels; 10 µl TEMED
for stacking gels)
Swirl gently to initiate polymerization.
Note: Prepare any desired volume of monomer solution by
using multiples of the 10 ml recipe. The volumes of APS and
TEMED must be adjusted accordingly.